Project 2: Serologic and molecular determinants of COVID-19 severity and immune protection

项目2摘要 项目2.新冠肺炎严重程度和免疫保护的血清学和分子决定因素。 关于SARS-CoV-2血清学的类型、动力学和持续时间，仍然存在悬而未决的科学问题 反应与患者的年龄、疾病严重程度、保护性免疫和传播的风险密切相关。 联系人。我们的总体目标是利用注释良好的STOP-COVID数据集和纵向样本 从第一反应者队列(有再次接触的高风险)和家庭接触者(项目1，核心B)收集到 进一步了解作为持久和保护性血清学基础的特定宿主和病毒因素 回应。我们的三个高度互动的目标包括：识别病毒中和抗体的差异 同型及其靶病毒蛋白/多肽/表位；评估针对疾病的交叉保护性免疫 通过对普通感冒冠状病毒的抗体加剧；并通过病毒宿主转录组分析，探索 SARS冠状病毒滴度或序列以及粘膜部位或血液的初始免疫反应与疾病相关 结果和上述血清学反应。目标1：确定免疫球蛋白亚型、效价、靶蛋白和 SARS-CoV-2特异性抗体的病毒中和活性及其与疾病严重程度和 保护：我们将使用VN检测(VNA)和荧光素酶免疫沉淀检测VN抗体 与病毒相关的同型特异性及其免疫优势和独特的靶病毒蛋白/肽/表位 疾病的严重性和防护。预期结果包括描绘暂时性抗体亚型的数据 和病毒靶标，以完善与保护性免疫相一致的血清学测试方案。目标2：确定 普通感冒冠状病毒和SARS-CoV-2特异性抗体对新冠肺炎防护和预防的影响 在急救人员及其家庭接触者的高暴露风险队列中的疾病严重性。诊断性 血清样本将用于针对独特和保守的CCCoV的创新ELISA平台 多肽和Spike假型CCCoV用于评估CCCoV抗体反应。预期结果包括 先前存在的冠状病毒抗体是否与新冠肺炎的临床症状、严重程度或 SARS-CoV-2感染的毒株特异性(目标3)，或再感染风险。目的3.粘膜和粘膜的相互关系模式 病毒和宿主遗传学的血清学免疫反应。我们将利用一种新型的靶向宿主-SARS-CoV-2 RNA测序试验(CoV-DXVX)以评估SARS-CoV-2 S1， S2、N蛋白或其他ORF影响疾病的表现、进展和预后。主机响应 通过粘膜或血液免疫基因诱导的概况和强度来衡量，将与 这些血清学反应和上述反应的持续时间、类型和保护能力。一旦疫苗被 部署到我们的第一反应者队列，所有3个目标将重点分析上述血清学反应 SARS-CoV-2血清阴性和阳性患者的免疫表型和宿主免疫基因诱导的差异 与自然感染的个体进行比较。

Project 2 Summary PROJECT 2. Serologic and molecular determinants of COVID-19 severity and immune protection. Unresolved scientific questions remain about how the type, kinetics and duration of SARS-CoV-2 serologic responses correlate with patient age, disease severity, protective immunity, and risk of spread to close contacts. Our overall goal is to utilize the well-annotated STOP-COVID data sets and longitudinal samples collected from the first-responder cohort (at high-risk for re-exposure) and family contacts (Project 1, Core B) to advance understanding of the specific host and viral factors that underlie long-lasting and protective serologic responses. Our three highly interactive aims include: to identify differences in virus neutralizing (VN) antibody isotypes and their target viral proteins/peptides/epitopes; to assess cross-protective immunity versus disease exacerbation by antibodies to common cold CoVs; and to probe, by virus-host transcriptome analysis, how SARS-CoV titer or sequence and initial immune response at mucosal sites or blood interrelate with disease outcomes and the above serologic responses. Aim 1: Define immunoglobulin isotypes, titer, target proteins and viral neutralization activity of SARS-CoV-2 specific Abs and their correlations with disease severity and protection: We will utilize VN assays (VNA) and a luciferase immunoprecipitation assay to identify VN antibody isotype specificities and their immunodominant and unique target viral proteins/peptides/epitopes relevant to disease severity and protection. Expected outcomes include data that delineate the temporal antibody isotypes and viral targets to refine protocols for serologic testing that align with protective immunity. Aim 2: Determine the impact of common cold CoVs (CCCoV) and SARS-CoV-2 specific antibodies on COVID-19 protection and disease severity in a high-exposure risk cohort of first responders and their household contacts. Diagnostic serum samples will be used in innovative ELISA platforms directed against unique and conserved CCCoV peptides, and Spike pseudotyped CCCoV to assess CCCoV antibody responses. Expected outcomes include original data on whether pre-existing CCCoV antibodies relate to COVID-19 clinical symptoms, severity or strain specificity of SARS-CoV-2 infection (Aim 3), or re-infection risk. Aim 3. Correlate patterns of mucosal and serologic immune responses with virus and host genetics. We will utilize a novel targeted host-SARS-CoV-2 RNA sequencing assay (CoV-DXVX) to assess whether virus titer or functional alterations in SARS-CoV-2 S1, S2, N proteins or other ORFs impact disease presentation, progression and outcome. Host response measured by the profile and strength of mucosal or blood immune gene induction will be correlated with the duration, type and protective ability of these serologic responses and with the ones above. Once vaccines are deployed to our first-responder cohort, all 3 aims will pivot to analyze how the above serologic response profiles and host immune gene induction differ in vaccinated SARS-CoV-2 seronegative and seropositive individuals compared with the naturally-infected individuals.